Return to Proposal Finder FY 2000 Proposal 20043

Proposal Table of Contents

Additional Documents

Section 1. General Administrative information
Section 2. Past accomplishments
Section 3. Relationships to other projects
Section 4. Objectives, tasks and schedules
Section 5. Budget
Section 6. References
Section 7. Abstract

Reviews and Recommendations
Title Type File Size File Date

Section 1. General Administrative Information

Title of Project Proposal Intracytoplasmic Sperm Injection: Genetic Retrieval From Single Sperm
BPA Project Proposal Number 20043
Business name of agency, institution,
or organization requesting funding
University of Idaho
Business acronym (if appropriate) UI

Proposal contact person or principal investigator

Name Joseph G. Cloud
Mailing Address
City, State, Zip Moscow, ID 83844-3051
Phone 2088856388
Fax 2088857905
Manager of program authorizing this project
Review Cycle FY 2000
Province Mainstem/Systemwide
Subbasin Systemwide
Short Description Develop methodology to retrieve the genetics from a single sperm by microinjection into an egg.
Target Species chinook salmon (Oncorhynchus tshawystcha) steelhead (Oncorhynchus mykiss)

Project Location

[No information]

Reasonable and Prudent Alternatives (RPAs)

Sponsor-Reported Relevant RPAs

Sponsor listed no RPAs for this project proposal

Relevant RPAs based upon NMFS & BPA Review

NMFS and BPA did not associate any reasonable and prudent alternatives with this project proposal

NPPC Program Measure Number(s) which this project addresses: 7.2, 7.4D, 7.4E (The 1994 Columbia River Basin Fish and Wildlife Program)
FWS/NMFS Biological Opinion Number(s) which this project addresses:
Other Planning Document References

CBFWA-Generated Information

Database Administrator notes on the history of this proposal form: None
Type of Project (assigned by CBFWA Analysts): anadromous

Section 2. Past Accomplishments

n/a or no information

Section 3. Relationships to Other Projects

Project ID Title Description Umbrella
9703800 Listed Salmonid Stocks Gamete Preservation ICSI of salmonid sperm will interact with the development of a sperm bank by ultimately providing insurance against accidental thawing of the cryopreserved materials No
Enhancement of Samonid Gamete Quality by Manipulation of Intracellualar ATP CO-PI; both projects have a goal of increasing the probability of retrieving the genetic information from sperm No
Endocrine Control of Ovarian Development in Salmonids A participant of the UI/WSU Fish Reproduction Program No
Analyzing Genetic and Behavioral Changes During Samonid Domestication A participant of the UI/WSU Fish Reproduction Program No
Induction of Precocious Sexual Maturity and Enhanced Egg Production in Fish A participant of the UI/WSU Fish Reproduction Program No
Viral Vaccines and Effects on Reproductive Status A participant of the UI/WSU Fish Reproduction Program No

Section 4. Objectives, Tasks and Schedules

Objectives and Tasks

Objective Task
1. Determine the pretreatment of sperm and the timing of the injection of the sperm relative to egg activation that maximizes male pronuclear development a. Describe the temporal changes in the decondensation of the sperm chromatin and the development of the male pronucleus following normal fertilization
1. b. Pretreat sperm with dithiothreitol and/or sonication before microinjection into the eggs. The measurable endpoint of this task is the proportion of eggs in which a male pronucleus is formed.
1. c. Inject the sperm into eggs at varying times following activation. The measurable endpoint will be the proportion of eggs in which a male pronuclues is formed.
2. Produce viable offspring from the injection of a single sperm into an activated egg a. Inject sperm with the gene for pigmentation into activatived, albino trout eggs (the protocol for sperm pretreatment and the time of injection will be dictated by the results of objective 1). The measurable endpoint will be normal, pigmented fry.
2. b. Inject steelhead sperm into activated steelhead eggs. The measurable endpoint will be normal, diploid fry.
2. c. Inject chinook sperm into activated chinook eggs. The measurable endpoint will be normal, diploid fry.

Objective Schedules and Costs

Objective Start Date End Date Measurable Biological Objectives Milestone FY 2000 Cost %
1 10/01/00 09/01/01 The proportion of eggs in which the male pronuclues has developed following the injection of a single sperm Producing a protocol for the pretreatment of the sperm and the timing of injection following egg activation for the microinjection of single sperm into salmonid eggs 38.0%
2 10/01/00 09/01/01 The proportion of normal, functional offspring produced by the intracytoplasmic injection of salmonid sperm into activated eggs The ability to produce offspring from a single sperm cell; the ability to retrieve the genetics from a single sperm cell; teach fisheries personel to microinject sperm into activated eggs. 62.0%

Section 5. Estimated Budget Summary

Itemized Budget

Item Note FY 2000 Cost
Personnel PI (4.0 mo); Scientific Aide (12 mo); Graduate Student (9.0 mo); Undergrad (320 hrs) $ 77,696
Fringe 28.5% for PI and Scientific Aide; 1.0% for students $ 16,940
Supplies $ 24,000
Operating equipment repair $ 4,240
Capital $ 8,765
Travel $ 3,000
Indirect $ 69,124
Other Administrative and Aquaculture Core Facilities $ 20,000
Total Itemized Budget $223,765

Total estimated budget

Total FY 2000 project cost $223,765
Amount anticipated from previously committed BPA Funds $ 0
Total FY 2000 budget request $223,765
FY 2000 forecast from 1999 $ 0
% change from forecast 0.0%

Reason for change in estimated budget

Not applicable

Reason for change in scope

Not applicable

Cost Sharing

Not applicable

Outyear Budget Totals

2001 2002 2003 2004
All Phases $234,953 $246,701 $259,036 $271,988
Total Outyear Budgets $234,953 $246,701 $259,036 $271,988

Other Budget Explanation

Schedule Constraints: None

Section 6. References

Reference Watershed?
Flahrety, S.P., D. Payne, N.J. Swann and C.D. Matthews. 1995. Assessmnt of fertilization failure and abnormal fertilization failure after intracytoplasmic sperm injection (ICSI). Reprod Fertil Dev 7:197-210. No
Gausen, D. 1993. The Norwegian gene bank programme for Atlantic Salmon. In: Cloud, J.G. and Throgaard, G.H. (eds) Genetic conservation of salmonid fishes. Plenum Press, New York pp181-188. No
Kimura, Y. and R. Yanagimachi. 1995. Intracytoplasmic sperm injection in the mouse. Biol Reprod 52:709-720. No
Palermo, G.D., H. Joris, P. Devroey and A.C. Van Strirteghem. 1992. Pregnancies after intracytoplasmic injection of a single spermatozoon into an oocyte. Lancet. 340:17-18. No
Tesarik, J. an M. Sousa. 1995. More than 90% fertilization rates after intracytoplasmic sperm injection and artifical induction of oocyte activation with calcium ionophore. Fertil Steril 63:343-349. No
Uehara, T. and R. Yanagimachi. 1976. Microsurigical injection of spermatozoa into hamster eggs with subsequent transformation of sperm nuclei into male pronuclei. Biol Reprod 15:467-470. No
Van Steirteghem, A.C., Z. Nagy, H. Joris, J. Liu, C. Straesen, J. Smitz, A. Wisamto and P. Devroey. 1993. High fertilization and implantation rates after intracytoplasmic sperm injection. Hum. Reprod. 8:1061-1066. No
Yanagida, K., R. Yanagimachi, S.D. Perreault and R.G. Kleinfeld. 1991. Thermostability of sperm nuclei assissed by microinjection into hamster oocytes. Biol Reprod. 44:440-447. No

Section 7. Abstract


The genetic contributions of male fish from threatened and endangered species are lost if their sperm cells are unable to fertilize eggs. A similar biological problem, human male factor infertility, exists in medicine. The human reproductive problem has been solved by the development of intracytoplasmic sperm injection (ICSI); normal human offspring are now routinely produced by the injection of a single sperm cell into an egg. Since the post-fertilization cellular events, sperm nuclear decondensation and the formation of the male pronucleus, appear to be very similar between mammals and fish, the overall objective of this proposed investigation is to produce fish by injecting a single sperm into an egg. The hypothesis to be tested is that the mechanical introduction of a sperm into the cytoplasm of an activated egg will result in the decondensation of the chromatin within the sperm nucleus, in the development of a morphologically normal male pronucleus, and in the production of a fully functional zygote. The experiments proposed for FY2000 are designed to develop the methodology of sperm intracytoplasmic injection for fish and to demonstrate that the DNA of the injected nucleus contributes to the genome of the resultant zygote. Ultimately, the ability to produce fish by ICSI is expected to provide the means to retrieve the genetics of sperm 1) from infertile or subfertile males, 2) that has been prematurely activated during collection, 3) that was cryopreserved and accidentally thawed, or 4) from dead or dying males.

Reviews and Recommendations

This information was not provided on the original proposals, but was generated during the review process.

This project has not yet been reviewed

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