# GENE EXPRESSION IN SINGLE AGING NEURONS AND GLIA

> **NIH NIH R01** · UNIVERSITY OF PENNSYLVANIA · 1991 · $166,373

## Abstract

In this proposal we examine the effects of aging on mRNA
expression in single neurons and glia isolated from the rat hippocampus.
A profile of gene expression in these cells will be generated by using
amplified antisense RNA (aRNA) made from single cells to probe Southern
blots containing the cloned cDNAs for a number of molecules thought to be
important in neuronal and glial cell development.  Characterization of
this profile for 50 individual neurons and glia will permit the
categorization of neurons and glia into groups dependent upon their
relative levels of expression of the selected mRNAs.  Generation of aRNA
probe from a single cell will be accomplished by injection of reverse
transcriptase and the oligo-dT-T7 amplification oligonucleotide directly
into dispersed calls using a patch-pipette.  Simultaneous with this
injection recordings will be taken from the cells to insure that they are
viable.  The cDNA made in this manner is of high complexity and is
converted into aRNA by making it double-stranded and. adding T7 RNA
polymerase to the template.  Several million-fold amplification of mRNA
levels have. been achieved using this technology. aRNA will be made from
several dispersed cells that are isolated from rats ranging in age from 2
days to 33 months.  These aRNAs will be used in the Southern blot
profiling screen and will also be cloned so that cDNA libraries from
single neurons and glia of these differing ages can be used to facilitate
isolation of previously uncharacterized mRNAs whose abundance is
age-related.  The glucocorticoid model of aging will be examined using
this same technology so that this model can be directly compared with
aging in the single cell.  Differences in age-related gene expression
between hippocampal tissue and single cells will be examined by comparing
the aRNA results with results generated using in situ transcription (1ST)
of hippocampal brain section.  Age-related and glucocorticoid regulated
mRNAs will be isolated by 1ST-based aRNA differential hybridization and
differential amplification.  Clones will be characterized by DNA
sequencing and in situ hybridization across the age range used to create
the single cell gene expression profile.

## Key facts

- **NIH application ID:** 3121840
- **Project number:** 1R01AG009900-01
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **SAM.gov UEI:** GM1XX56LEP58
- **Principal Investigator:** JAMES H EBERWINE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1991
- **Award amount:** $166,373
- **Award type:** 1
- **Project period:** 1991-05-03 → 1996-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3121840

## Citation

> US National Institutes of Health, RePORTER application 3121840, GENE EXPRESSION IN SINGLE AGING NEURONS AND GLIA (1R01AG009900-01). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3121840. Licensed CC0.

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