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GRANT WRITING AND BUDGETING. Marc E. Freeman, Ph.D. Department of Biological Science Florida State University Tallahassee, FL 32306 850 644-3896. HOW TO WRITE YOUR FIRST GRANT. Nothing beats a good idea Be realistic Make the presentation clear and simple

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grant writing and budgeting

GRANT WRITING AND BUDGETING

Marc E. Freeman, Ph.D.

Department of Biological Science

Florida State University

Tallahassee, FL 32306

850 644-3896

how to write your first grant
HOW TO WRITE YOUR FIRST GRANT
  • Nothing beats a good idea
  • Be realistic
  • Make the presentation clear and simple
  • Make the presentation easy to read
  • Present yourself as the greatest expert in the field
  • Submit a realistic budget
nothing beats a good idea
NOTHINGBEATS A GOOD IDEA
  • Articulate a worthwhile, single, focused objective
  • Articulate Specific Aims that are clearly related to one another and logically fit under the umbrella of the overall objective
  • Present gaps in our knowledge
  • Plant the seed for achieving each specific aim by presenting the questions to be asked which will fill the gaps
be realistic
BE REALISTIC
  • Ask questions which are answerable
  • Provide tantalizing preliminary data as evidence that the questions are worth asking and answerable
  • Propose technical approaches which are within the realm of your published technical expertise OR provide preliminary data
  • The volume of work proposed should be proportional to the time of support requested and your other obligations
make the presentation clear and simple
MAKE THE PRESENTATION CLEAR AND SIMPLE
  • Assume total ignorance on the part of the reviewer
  • Provide all of the simplest conceptual background
  • No abbreviations or acronyms without definition
  • Use diagrams and cartoons to illustrate concepts
  • Use formatting for emphasis
  • Be redundant
make the presentation easy to read
MAKE THE PRESENTATION EASY TO READ
  • Think of the reviewer
  • Avoid verbosity
  • Adopt a unique style of presentation
  • Tell the reviewer what he is supposed to think and write
  • Do not force the reviewer to hunt through the application for information
present yourself as the greatest expert in the field
PRESENT YOURSELF AS THE GREATEST EXPERT IN THE FIELD
  • Know the literature in depth and breadth
  • Do not make statements without attribution or preliminary data
  • Do not be reluctant to admit shortcomings
  • Seek collaborators or mentors when your expertise cannot be documented
submit a realistic budget
SUBMIT A REALISTIC BUDGET
  • Request only what you need and you can defend
  • Justify every item in the budget thoroughly
  • Do not request less than you need
  • Present evidence that your institution supports your research
the research plan
THE RESEARCH PLAN
  • Specific Aims
  • Background and Significance
  • Preliminary Studies
  • Research Design and Methods
  • Human Subjects
  • Vertebrate Animals
  • Literature Cited
  • Consortium/Contractual Arrangements
  • Consultants
specific aims
SPECIFIC AIMS
  • One short paragraph to identify system
  • One short paragraph to describe regulatory steps of the system
  • One short paragraph containing a one sentence CENTRAL HYPOTHESIS
  • One short sentence stating each aim individually
  • Follow each with a simple statement describing what you will do to satisfy that aim
slide11

Research Plan

Specific Aims

Prolactin (PRL) is one of the most versatile hormones of mammalian organisms. Besides its role in lactation, secretion of PRL contributes to a wide range of physiological functions, i.e. adaptation to new environment (22), immune functions (23) osmoregulation (24,25), reproduction (26) and behavior (27). PRL of anterior pituitary origin is secreted by lactotrophs (28). Lactotrophs have spontaneously high secretory activity, which is controlled primarily by tonic inhibitory input of hypothalamic origin (29), though they receive stimulatory input, as well (29-31).

The physiological PRL inhibiting factor is dopamine (DA) (32,33). DA is released from three, anatomically and functionally distinct hypothalamic neuroendocrine cell populations: I). periventricular hypothalamic dopaminergic (PHDA) neurons of the periventricular nucleus (A14), II) tuberohypophysial (THDA) and III) tuberoinfundibular (TIDA) neurons (34) of the arcuate nucleus (A12). PHDA and THDA neurons terminate in the intermediate and neural lobes of the pituitary gland, respectively (35,36). DA, released from THDA/PHDA terminals may reach the anterior lobe through the short portal vessels. On the other hand, TIDA neurons terminate in the external zone of the median eminence supplying the anterior pituitary gland with DA through the long portal vessels. Although the central role of TIDA neurons in hypothalamic control of PRL secretion is acknowledged, the relative contribution and importance of the distinct neuroendocrine DAergic neuron subpopulations to the regulation of PRL secretion is not understood.

slide12
Studies from our (37-39) and other (40-43) laboratories suggest that the daily activities of all three DAergic neuroendocrine neuron populations exhibit similar rhythmic changes (i.e. high in the morning, low in the afternoon). Although, the amplitudes of these rhythms of TIDA, THDA and PHDA populations are differentially modulated by ovarian steroids, the presence of rhythms is independent of the ovarian steroid background in female rats in all neuroendocrine DAergic neuron populations. Our CENTRAL HYPOTHESIS is that the rhythmic activity in all three populations of neuroendocrine DAergic neurons are paced by direct circadian input from the suprachiasmatic nucleus while its amplitude is modulated by interneuron-mediated input, ovarian steroid background, and PRL feedback from the pituitary gland. In this application, we will focus on the role of direct suprachiasmatic input, PRL feedback and signaling mechanisms in the generation of rhythmic patterns of DA release from the TIDA, THDA and PHDA by pursuing the following SPECIFIC AIMS:
slide13

To characterize the chronobiological and anatomical basis of regulatory mechanisms governing the rhythmic patterns of the DAergic input on PRL secretion. By conducting in vivo experiments in constant environments we will test whether the daily changes in the activities of neuroendocrine DAergic neuron populations are light entrained endogenous circadian rhythms. Tract tracing studies will be used to investigate the functional anatomical nature of the direct connection(s) between the circadian zeitgeber in the suprachiasmatic nucleus and the different neuroendocrine DAergic subpopulations. Antisense antagonism will be used to verify the functional significance of suprachiasmatic efferent fibers on the neuroendocrine DAergic neuron populations.

To characterize the feedback role of PRL in the daily activity of PHDA, THDA and TIDA neurons. The incidence of PRL-Rs in the THDA and TIDA neurons is higher than in PHDA neurons, although PHDA neurons also express PRL-R in ovarian steroid-replaced rats. We suggest that PRL feedback is inherently part of the THDA and TIDA neurons’ circadian regulation and it is inducible in PHDA neurons by ovarian steroids. In order to verify this hypothesis, we will compare the effects of PRL on the activity of PHDA, THDA and TIDA neuron populations in ovariectomized and ovarian steroid-replaced rats.

To identify the signal transduction mechanisms involved in the circadian regulation of the secretory activity of hypothalamic neuroendocrine DAergic neurons. We will identify the receptors, second messenger and cellular effector systems involved in mediating the effects of circadian input and PRL feedback on the neurosecretory activity of the PHDA, THDA and TIDA neuron subpopulations.

slide14
BACKGROUND AND SIGNIFICANCE
  • Provide just enough background information so the reviewer appreciates what you are proposing
  • Extraneous information is distracting
  • Compartmentalize information with bold headings, key words and sentences
  • Make copious use of diagrams and cartoons
  • Use a terminal sentence pointing to your goal at the end of each compartment
slide15

Fig. 1

The neuroendocrine DAergic neuron populations and their feedback regulation (Fig. 1)

The hypothalamic A14 and A12 cell groups (Fig. 1), which are exclusively comprised of DAergic neurons (64,65) provide the pituitary gland with DA through two different routes: I) The tuberoinfundibular DAergic (TIDA) neurons send short projections to the external zone (EZ) of the median eminence. TIDA axons terminate on the basement membrane of the perivascular space surrounding the primary capillary loops of the portal system. From here the long portal vessels (LP) carry DA of TIDA origin to the anterior lobe (AL) of the pituitary gland. Perikarya of TIDA neurons show bimodal rostrocaudal distribution (66). The majority of TIDA neurons originate throughout the arcuate nucleus (ARN, A12), and a smaller

population arises from the periventricular nucleus (PeVN, A14) (66). II) The tuberohypophysial DAergic (THDA) neurons of the rostral ARN (A12) and the periventricular hypothalamic (PHDA) neurons of the PeVN (A14) have long axons, which course through the pituitary stalk (PS) and terminate in the neural (NL) and intermediate(IL) lobes of the pituitary gland, respectively (35). Short portal vessels(SP) provide communication between the neurointermediate and the AL (Figure 1).

slide16
The terminal areas of the TIDA, THDA and PHDA axons are neurohemal zones, which lack the blood-brain barrier. The absence of the blood-brain barrier and the continuity between the pericapillary space and the liquor space are the anatomical bases for a free, most likely bi-directional communication between axon terminals and the blood-liquor space (67). While all three neuroendocrine neuron populations express PRL-Rs in female rats, the incidence of PRL-Rs on THDA and TIDA neurons is higher than in PHDA neurons (21). Changes in the ovarian steroid background and/or serum PRL levels differentially affect the abundance of PRL-R in the neuroendocrine DAergic neuron populations (21). Although to different extents, the activities ofTIDA (68,69), THDA and PHDA neurons are indeed affected by PRL in the peripheral blood (DeMaria, Lerant and Freeman, 1999, submitted). Of these neuroendocrine DAergic neuron populations, the autoregulatory properties of the PHDA neurons seem to most closely resemble the well-characterized nigrostriatal dopaminergic neurons as both nigrostriatal DAergic (70,71) and PHDA neurons (72) seem to be inhibited by D2/3–type DA (auto)receptors. There are data, however, indicating that TIDA neurons can be influenced by both D1 and D2 receptors, but the responses are different from that seen in nigrostriatal DAergic neurons (73). D2 receptor agonists were reported to stimulate TIDA neurons (74), while specific D1 agonists inhibit the reserpine- or neurotensin-evoked TIDA activation (75). We provide preliminary data demonstrating that D2 receptors do not colocalize with TIDA neuron terminals in the median eminence. This suggests that D2/3 receptors do not act as presynaptic autoreceptors on TIDA neurons. We will test our hypothesis that PRL provides the primary feedback in the THDA and TIDA neurons instead of D2/3 autoreceptors.
significance
SIGNIFICANCE
  • Be imaginative
  • Avoid unrealistic ideas
  • Keep it simple
  • Be brief
  • Present in layman’s terms if possible
preliminary experiments
PRELIMINARY EXPERIMENTS
  • Present preliminary data which supports the feasibility of each specific aim
  • Do not present preliminary data which will completely accomplish the specific aim
  • Present preliminary data which proves you can use a new technique
  • Present preliminary data to support your development and validation of a previously undescribed technique
research design and methods general methods
RESEARCH DESIGN AND METHODSGENERAL METHODS
  • Present most often used methods first each in separate titled paragraphs
  • Present in a depth which is inversely proportional to your published experience with the methods
  • Cite publications in which you have used the methods
  • Refer to the preliminary data when describing unpublished methods
slide20
Preparation of tissue for immunocytochemistry (ICC)

Rats are sacrificed by an overdose of sodium pentobarbital, and perfused through a transcardial cannula with 50 ml of ice cold phosphate buffer (PBS, 0.1 M, pH 7.34, 295 mosm) immediately followed by 100 ml of ice cold fixative. The fixative applied usually is 4 % paraformaldehyde (PFA, in 0.1 M PBS, pH 7.5). The brains and pituitaries are postfixed for an hour in situ. After removal the brains and pituitaries are rinsed in PBS and immersed in 20% sucrose solution (in PBS, at 4ºC) until sinking. The tissue blocks are frozen on the freezing stage of a HM500OM cryostat (Zeiss, Germany). The brains are cut in 35-m coronal section between 300-4200 m post bregma and collected into 4 parallel series of free-floating sections. The sections are stored in cryoprotectant solution (133) until ICC is initiated. If the aforementioned tissue block does not contain an area with established immunoreactivity for the antigen in question, other areas of the brain (or other organs) are processed to provide positive and negative controls for the immunostaining. The pituitary glands are cut in 10-20 m coronal or horizontal sections, thaw-mounted on gelatin-subbed glass microscope slides and stored at -80ºC in closed tissue boxes containing Dryerite. Both the free floating brain sections and the mounted pituitary sections are thoroughly washed (5 X in PBS) prior to ICC. We have had ample experience with these techniques (21,37,38)

research design and methods experimental approaches to each specific aim
RESEARCH DESIGN AND METHODSExperimental approaches to each Specific Aim
  • Each specific aim is its own section
  • State specific aim
  • State hypothesis associated with that specific aim
  • State question(s) associated with that hypothesis
  • Provide rationale for each question
  • Describe experiments
  • Expected results, interpretation, shortcomings and pitfalls
outline of a typical specific aim
OUTLINE OF A TYPICAL SPECIFIC AIM
  • Specific Aim I

Hypothesis A

Basis of hypothesis A

Question 1

Rationale for question

Experiment 1, 2, 3, 4……….

Question 2

Rationale for question

Experiment 1, 2, 3, 4……….

Experimental results, interpretation, shortcomings, pitfalls

Hypothesis B………….

hypothesis
HYPOTHESIS
  • Each Specific Aim should be HYPOTHESIS DRIVEN

May consist of more than one hypothesis

  • Hypothesis should be defended in terms of the overall objectives previously stated

Do not be afraid of redundancy with the Background

experimental questions
EXPERIMENTAL QUESTIONS
  • Each question should logically fit under the umbrella of its hypothesis
  • The rationale for each question should serve to clarify the question, the necessity for asking the question and its relationship to the hypothesis

What is to be gained by asking the question?

experiments
EXPERIMENTS
  • Describe each experiment in sufficient detail so the reviewer can understand it and believe you can successfully complete it

Do NOT forget controls

Do NOT forget statistical analyses

Explain rationale for doses and times

Refer to previously described general methods

Describe methods unique to this experiment

expected results and interpretation
EXPECTED RESULTS AND INTERPRETATION
  • State ALL expected and unexpected results
  • State the interpretation of each

What will you do if one of the unanticipated results emerge?

Be very careful that an unanticipated result does not doom the subsequent specific aims/questions

shortcomings pitfalls limitations
SHORTCOMINGS, PITFALLS, LIMITATIONS
  • What are possible alternative approaches?

Why are you using the approach you are?

Why are you not using one of the alternatives?

What are the strengths and weaknesses of the approach you are using?

  • Are there any pitfalls you foresee?

How will you deal with them?

  • What are your lab’s strengths and weaknesses?
human subjects vertebrate animals
HUMAN SUBJECTS/VERTEBRATE ANIMALS
  • Follow the instructions carefully
  • Verbosity with detail is good
  • Be realistic about number of subjects/animals
  • Be aware of rules governing limitations of use
summary statement
SUMMARY STATEMENT
  • I assist the reviewer by writing my own summary statement as if I were the reviewer

Significance, Approach, Innovation, Investigator, Environment

  • Be brutally honest

Strengths and Weaknesses

Shortcomings

  • What will the project accomplish?
  • What will the project not accomplish?
consultants collaborators
CONSULTANTS/COLLABORATORS
  • Use consultants for techniques that you intend to use for the first time

Secure letters confirming their role

  • Interest collaborators in supplying unique reagents that are critical to your work

Antibodies, nucleotides, peptides

Obtain letters of collaboration

  • Have a senior level colleague with the greatest experience obtaining grants and serving on Study Sections read and critique the application
you can control the fate of the application before mailing
YOU CAN CONTROL THE FATE OF THE APPLICATION BEFORE MAILING
  • Review the charge and membership of the possible Study Sections to which it may be assigned

http://www.csr.nih.gov/committees/rosterindex.asp#A

  • Write a cover letter to be included in your application package

Request assignment to the Study Section you select

common reasons grant applications receive unfundable scores
COMMON REASONS GRANT APPLICATIONS RECEIVE UNFUNDABLE SCORES
  • Poor organization

Not an integrated body of work

  • Exercise in data collection

“Fishing expedition”

  • Work too descriptive and not experimental

“OK, so you showed that there is message for the insulin receptor on the pancreas, now what?”

common reasons grant applications receive unfundable scores33
COMMON REASONS GRANT APPLICATIONS RECEIVE UNFUNDABLE SCORES
  • Lack of sufficient detail
  • Insufficient convincing preliminary data
  • Applicant not capable of performing the work
  • Inadequate institutional support
  • Objective not very important to health and disease
  • Overly ambitious
what do i do if application is not approved for grant funding
WHAT DO I DO IF APPLICATION IS NOT APPROVED FOR GRANT FUNDING?
  • Find out why

Ask SRA and Institute official

Obtain Summary Statement

Read Summary Statement thoroughly and often

Ask a mentor to read Summary Statement

Mentor and yourself should highlight issues for attention

what do i do if application is not approved for grant funding35
WHAT DO I DO IF APPLICATION IS NOT APPROVED FOR GRANT FUNDING?
  • Re-write application
  • Respond to Summary Statement by:

Enumerating each criticism and the actions you propose to take

what actions should i take
WHAT ACTIONS SHOULD I TAKE?
  • Take the criticism constructively

No finger pointing or accusations

Admit the reviewers were right

  • Articulate your response to each

Reorganize and simplify presentation

Provide more detail

Provide preliminary data in response

Get a consultant

Add experiments

Eliminate experiments

DIPLOMATICALLY point out reviewer error

slide37
This is a revision of our previous application reviewed by the Biochemical Endocrinology (BCE) Study Section in October 1998. We have taken the reviewers’ critiques constructively and made the requested changes, which we believe, improved the current application. We thank the reviewers for providing insight into the shortcomings of our proposal. We have addressed each criticism below and have made appropriate changes in the text as indicated in Century Schoolbook font.
  • Response to Critique 1:
  • Criticism: The second question posed in Specific Aim I does not relate the changes in neuronal activity associated with the rhythm of water intake to the pharmacology of the DA receptor under investigation and no discussion of the effects of dehydration on food intake and other variables is presented.
  • Response: In agreement with the reviewer, we have eliminated the water deprivation experiments from this proposal.
slide38
Criticism: The rationale for use of unilateral versus bilateral injection sites of [biotinylated dextran] tracer is not apparent nor is the choice of a seven day waiting period following tracer injection justified.
  • Response: To answer the question of anatomical projection, unilateral injections will provide sufficient information. Also, it is easier to interpret the results of unilateral injections, since we cannot exclude the possibility of SCN efferents crossing the midline. The seven day survival period gives excellent results with the BDA-F tracer as shown by several laboratories (1-3). We now include these explanations and citations at the proposed experiments as well.
and now
AND NOW………….
  • You can do nothing more…….

SO……

AWAIT THE AWARD NOTICE!!!!!!!