Activation of gene expression of the gamma-aminobutyric acid rather than the glutamatergic system in the preoptic area during the preovulatory gonadotropin surge of the rat.
There is increasing evidence that in the rat prior to and during the preovulatory LH surge, release rates of GABA in the preoptic area (POA) are decreased while no such changes occurred in the mediobasal hypothalamus (MBH). In addition, GnRH release appears to be facilitated by an increased preoptic excitation of glutamate (GLU). To investigate whether such changes of secretory activity of intrahypothalamic GABA or GLU neurons are associated with altered gene expression of biosynthetic enzymes or transporter proteins characteristic for either neuronal system, we determined mRNA levels of the two forms of the GABA-synthesizing enzyme glutamate decarboxylase (GAD65 and GAD67), the glutamate-synthesizing enzyme glutaminase (GLS), the GABA transporter type 1 (GAT-1) and the glutamate-aspartate transporter type 1 (GLAST). Competitive RT-PCRs using mutant cRNAs as internal standards were conducted with mRNA extracted from microdissected tissue of POA and MBH from diestrous, proestrous, and estrous rats. Proestrous animals were subgrouped according to their endocrine status as follows: 'prior to', on the 'ascending' or on the 'descending' limb of the LH peak, and 'after the LH surge (post)'. During the preovulatory LH surge, mRNA concentrations of GAD67 and GAT-1 in the POA were significantly increased compared to those observed on diestrous (2.8-fold for GAD67 and 2.5-fold for GAT-1, p < 0.01), while in the MBH the amount of both mRNAs remained constant. The expression levels of GAD65, GLS and GLAST were without any changes in the POA as well as in the MBH. These findings support the hypothesis that in rats induction of the preovulatory LH surge is controlled at the level of GnRH perikarya, and suggest that altered activities of intrapreoptic GABA neurons at both transcriptional and secretory levels are pivotal for the preovulatory activation of GnRH neurons.
Leonhardt
S
S
Division of Clinical Endocrinology, Department of Obstetrics and Gynecology, University of Göttingen, Germany.
Böning
B
B
Luft
H
H
Wuttke
W
W
Jarry
H
H
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D027322:Amino Acid Transport System X-AG; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D004958:Estradiol / Q000097:blood; D005260:Female; D050485:GABA Plasma Membrane Transport Proteins; D015870:Gene Expression / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005968:Glutamate Decarboxylase / Q000235:genetics*; D018698:Glutamic Acid / Q000378:metabolism*; D005972:Glutaminase / Q000235:genetics; D007986:Luteinizing Hormone / Q000097:blood; D008565:Membrane Proteins / Q000235:genetics*; D026901:Membrane Transport Proteins; D009474:Neurons / Q000737:chemistry / Q000201:enzymology; D027361:Organic Anion Transporters; D011301:Preoptic Area / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D011359:Proestrus / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D018528:ATP-Binding Cassette Transporters; D027322:Amino Acid Transport System X-AG; D002352:Carrier Proteins; D050485:GABA Plasma Membrane Transport Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D027361:Organic Anion Transporters; D012333:RNA, Messenger; C493415:Slc6a1 protein, rat; D018698:Glutamic Acid; D004958:Estradiol; D005680:gamma-Aminobutyric Acid; D007986:Luteinizing Hormone; D005972:Glutaminase; D005968:Glutamate Decarboxylase