國立虎尾科技大學 |

Functional Analysis of the Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (GABRA1) Gene During Zebrafish Development.

Record Type:	Language materials, manuscript : Monograph/item
Title/Author:	Functional Analysis of the Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (GABRA1) Gene During Zebrafish Development./
Author:	Reyes-Nava, Nayeli Gabriela.
Description:	1 online resource (179 pages)
Notes:	Source: Dissertations Abstracts International, Volume: 85-03, Section: B.
Contained By:	Dissertations Abstracts International85-03B.
Subject:	Developmental biology. -
Online resource:	click for full text (PQDT)
ISBN:	9798380178372

Functional Analysis of the Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (GABRA1) Gene During Zebrafish Development.
Reyes-Nava, Nayeli Gabriela.

Functional Analysis of the Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (GABRA1) Gene During Zebrafish Development. - 1 online resource (179 pages)

Source: Dissertations Abstracts International, Volume: 85-03, Section: B.

Thesis (Ph.D.)--The University of Texas at El Paso, 2023.

Includes bibliographical references

The GABRA1 gene encodes for the alpha-1 (α1) subunit of the Gamma-Aminobutyric acid type A receptor (GABAAR), which are the primary modulators of synaptic inhibition in the central nervous system (CNS). Alpha-1 subunits are essential for maintaining the normal function of native receptors and contribute to over 60% of all GABAARs in the CNS. Remarkably, a broad spectrum of neurodevelopmental and epilepsy-associated disorders have been linked with mutations in the GABRA1 gene. However, the developmental, behavioral, and molecular mechanisms underlying GABRA1-associated epileptic disorders remain to be fully understood. Hence, the overarching goal of this dissertation is to investigate the behavioral and molecular mechanisms associated with GABRA1 deficiency during brain development. Two independent experimental approaches were taken using zebrafish as a model system to address this goal. Our first study describes the identification of a de novo missense variant of GABRA1 and the in vivo functional analysis of gabra1 using morpholino-mediated knockdown. We developed a behavioral paradigm using the Zebrabox technology to investigate the behavioral consequences of gabra1 loss of function. Interestingly, transient knockdown of gabra1 resulted in hypomotility at early larval stages. This hypomotile phenotype was correlated with altered expression of genes that encode for other GABAAR subunits. Mainly we show downregulation of the β2 and γ2 transcripts, which are the primary assembly partners of α1, and upregulation of the homologous α6b subunit. Despite these molecular defects in the expression of major GABAAR subunits, morphant larvae exhibited a positive response to a potent GABAAR antagonist, pentylenetetrazol (PTZ), indicating the presence of an active receptor despite the deficient expression of gabra1. Our findings contrast with those described in a zebrafish germline mutant of gabra1 generated previously. The distinct behavioral phenotypes between knockdown and germline mutation of gabra1 prompted us to characterize an additional gabra1 germline mutant. Therefore, our second unpublished study describes the behavioral and molecular analysis of an uncharacterized nonsense mutant of gabra1 (sa43718 allele) generated by the Wellcome Sanger Institute as part of the zebrafish mutation project in 2016. Behavioral analysis of this allele resulted in lightinduced hyperactive locomotion resembling generalized seizures. Targeted mRNA analysis revealed that this behavioral phenotype was associated with reduced expression of gabra1 and upregulation of gabra4, which encodes the a4 subunit. Mutant larvae were treated with PTZ to examine whether gabra1 mutants had an active receptor. Notably, mutant larvae exhibited reduced response compared to controls, suggesting that a GABAAR with reduced functionality is present. Subsequent proteomic analysis of gabra1 mutants revealed abnormal expression of proteins essential for synaptic vesicle transport, mitochondrial function, and potassium and sodium homeostasis regulation. Collectively, we highlight the essential role of gabra1 in early development and epileptic phenotypes.

Electronic reproduction.
Ann Arbor, Mich. :
ProQuest,
2024

Mode of access: World Wide Web

ISBN: 9798380178372Subjects--Topical Terms:

669036
Developmental biology.
Subjects--Index Terms:

Behavior mechanismsIndex Terms--Genre/Form:

554714
Electronic books.

Functional Analysis of the Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (GABRA1) Gene During Zebrafish Development.
LDR:04621ntm a2200409K 4500 001 1141467
005 20240315085549.5
006 m o d
007 cr mn ---uuuuu
008 250605s2023 xx obm 000 0 eng d
020 $a 9798380178372
035 $a (MiAaPQ)AAI30634678
035 $a AAI30634678
040 $a MiAaPQ $b eng $c MiAaPQ $d NTU
100 1 $a Reyes-Nava, Nayeli Gabriela. $3 1465247
245 1 0 $a Functional Analysis of the Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (GABRA1) Gene During Zebrafish Development.
264 0 $c 2023
300 $a 1 online resource (179 pages)
336 $a text $b txt $2 rdacontent
337 $a computer $b c $2 rdamedia
338 $a online resource $b cr $2 rdacarrier
500 $a Source: Dissertations Abstracts International, Volume: 85-03, Section: B.
500 $a Advisor: Quintana, Anita M.
502 $a Thesis (Ph.D.)--The University of Texas at El Paso, 2023.
504 $a Includes bibliographical references
520 $a The GABRA1 gene encodes for the alpha-1 (α1) subunit of the Gamma-Aminobutyric acid type A receptor (GABAAR), which are the primary modulators of synaptic inhibition in the central nervous system (CNS). Alpha-1 subunits are essential for maintaining the normal function of native receptors and contribute to over 60% of all GABAARs in the CNS. Remarkably, a broad spectrum of neurodevelopmental and epilepsy-associated disorders have been linked with mutations in the GABRA1 gene. However, the developmental, behavioral, and molecular mechanisms underlying GABRA1-associated epileptic disorders remain to be fully understood. Hence, the overarching goal of this dissertation is to investigate the behavioral and molecular mechanisms associated with GABRA1 deficiency during brain development. Two independent experimental approaches were taken using zebrafish as a model system to address this goal. Our first study describes the identification of a de novo missense variant of GABRA1 and the in vivo functional analysis of gabra1 using morpholino-mediated knockdown. We developed a behavioral paradigm using the Zebrabox technology to investigate the behavioral consequences of gabra1 loss of function. Interestingly, transient knockdown of gabra1 resulted in hypomotility at early larval stages. This hypomotile phenotype was correlated with altered expression of genes that encode for other GABAAR subunits. Mainly we show downregulation of the β2 and γ2 transcripts, which are the primary assembly partners of α1, and upregulation of the homologous α6b subunit. Despite these molecular defects in the expression of major GABAAR subunits, morphant larvae exhibited a positive response to a potent GABAAR antagonist, pentylenetetrazol (PTZ), indicating the presence of an active receptor despite the deficient expression of gabra1. Our findings contrast with those described in a zebrafish germline mutant of gabra1 generated previously. The distinct behavioral phenotypes between knockdown and germline mutation of gabra1 prompted us to characterize an additional gabra1 germline mutant. Therefore, our second unpublished study describes the behavioral and molecular analysis of an uncharacterized nonsense mutant of gabra1 (sa43718 allele) generated by the Wellcome Sanger Institute as part of the zebrafish mutation project in 2016. Behavioral analysis of this allele resulted in lightinduced hyperactive locomotion resembling generalized seizures. Targeted mRNA analysis revealed that this behavioral phenotype was associated with reduced expression of gabra1 and upregulation of gabra4, which encodes the a4 subunit. Mutant larvae were treated with PTZ to examine whether gabra1 mutants had an active receptor. Notably, mutant larvae exhibited reduced response compared to controls, suggesting that a GABAAR with reduced functionality is present. Subsequent proteomic analysis of gabra1 mutants revealed abnormal expression of proteins essential for synaptic vesicle transport, mitochondrial function, and potassium and sodium homeostasis regulation. Collectively, we highlight the essential role of gabra1 in early development and epileptic phenotypes.
533 $a Electronic reproduction. $b Ann Arbor, Mich. : $c ProQuest, $d 2024
538 $a Mode of access: World Wide Web
650 4 $a Developmental biology. $3 669036
650 4 $a Molecular biology. $3 583443
650 4 $a Biology. $3 599573
650 4 $a Aquatic sciences. $3 1178821
653 $a Behavior mechanisms
653 $a Epilepsy
653 $a Central nervous system
653 $a Pentylenetetrazol
653 $a Zebrafish
655 7 $a Electronic books. $2 local $3 554714
690 $a 0758
690 $a 0307
690 $a 0306
690 $a 0792
710 2 $a ProQuest Information and Learning Co. $3 1178819
710 2 $a The University of Texas at El Paso. $b Biological Sciences. $3 1465248
773 0 $t Dissertations Abstracts International $g 85-03B.
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30634678 $z click for full text (PQDT)