Immunoreactivity to neurofilaments in the rodent anterior pituitary is associated with the expression of alpha 1A protein subunits of voltage-gated Ca2+ channels.
We recently reported that rodent anterior pituitary (AP) cells (with the exception of corticotrophs and melanotrophs) express neuronal markers, including 68-kDa neurofilaments (NF68) in an oestrogen-dependent manner. The functional significance of neurofilament (NF) expression in the AP is unknown, but recent data in myelinated nerve fibres from NF-null mice suggest that NFs can regulate ion channel function. Because Ca(2+) influx through voltage-gated Ca(2+) channels is required for hormone secretion in AP cells, and oestrogen regulates the expression of Ca(2+) channels in AP cells, the present study examined the expression of alpha1 subunits of voltage gated Ca(2+) channels in relation to that of NF68. Using quantitative immunofluorescence, we demonstrate that alpha 1C and alpha 1D subunits are abundantly expressed in female AP cells, alpha 1A subunits are moderately expressed, and alpha 1G and alpha 1B subunits are expressed at the lowest levels. Double-immunostaining showed that NF68 expression is not correlated with that of alpha 1C, alpha 1D or alpha 1B. Expression of alpha 1G and NF68 appear to be mutually exclusive from each other. Moreover, alpha 1A subunit and NF68 expression are significantly correlated and alpha 1A immunoreactivity is sexually dimorphic (i.e. low in males and high in females) and its levels of expression vary during the oestrous cycle, similar to NF68. Finally, omega-agatoxin IVA, a specific blocker of P/Q type Ca(2+) currents that are a result of the activity of alpha 1A subunits, inhibited to a greater extent spontaneous [Ca(2+)](i) fluctuations in AP cells from females in oestrous and dioestrous, whereas cells from females in pro-oestrous and males were less affected by this toxin. These results suggest a preferential participation of P/Q-type Ca(2+) channels and hence alpha 1A subunits, in regulating spontaneous Ca(2+) transients in AP cells under conditions where the proportion of NF68-expressing cells is high. It remains to be determined whether the expression of NF68 affects that of alpha 1A Ca(2+) channel subunits or vice versa.
Fiordelisio
T
T
Instituto de Fisiología Celular, Departamento de Biofísica, Universidad Nacional Autónoma de México, Circuito Exterior, Ciudad Universitaria, México City, Mexico. tfiorde@ifc.unam.mx
Jiménez
N
N
Baba
S
S
Shiba
K
K
Hernández-Cruz
A
A
D000818:Animals; D002118:Calcium / Q000378:metabolism; D020864:Calcium Channels, N-Type; D020865:Calcium Channels, P-Type / Q000235:genetics / Q000378:metabolism*; D020907:Calcium Channels, Q-Type / Q000235:genetics / Q000378:metabolism*; D030762:Estrous Cycle / Q000502:physiology; D005260:Female; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*
D002118:Calcium; D020864:Calcium Channels, N-Type; D020865:Calcium Channels, P-Type; D020907:Calcium Channels, Q-Type; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D021122:Protein Subunits; C044882:neurofilament protein L