The origins of the circumventricular organs.

The circumventricular organs (CVOs) are specialised neuroepithelial structures found in the midline of the brain, grouped around the third and fourth ventricles. They mediate the communication between the brain and the periphery by performing sensory and secretory roles, facilitated by increased vascularisation and the absence of a blood-brain barrier. Surprisingly little is known about the origins of the CVOs (both developmental and evolutionary), but their functional and organisational similarities raise the question of the extent of their relationship. Here, I review our current knowledge of the embryonic development of the seven major CVOs (area postrema, median eminence, neurohypophysis, organum vasculosum of the lamina terminalis, pineal organ, subcommissural organ, subfornical organ) in embryos of different vertebrate species. Although there are conspicuous similarities between subsets of CVOs, no unifying feature characteristic of their development has been identified. Cross-species comparisons suggest that CVOs also display a high degree of evolutionary flexibility. Thus, the term 'CVO' is merely a functional definition, and features shared by multiple CVOs may be the result of homoplasy rather than ontogenetic or phylogenetic relationships.

Central cholinergic mechanisms mediate swallowing, renal excretion, and c-fos expression in the ovine fetus near term.

Fetal swallowing and renal metabolism contribute importantly to amniotic and body fluid homeostasis. To determine central cholinergic modulation of swallowing activity and renal excretion associated with neural activity, we examined the effects of intracerebroventricular injection of carbachol, a cholinergic agonist, in ovine fetuses at 0.9 gestation. Fetuses were chronically prepared with thyrohyoid, nuchal and thoracic esophagus, and diaphragm electromyogram electrodes, as well as lateral ventricle and vascular catheters. Electrodes were also implanted on the parietal dura for determination of fetal electrocorticogram (ECoG). After 5 days of recovery, fetal swallowing, ECoG, and urine output were monitored during basal period and the experimental period following intracerebroventricular injection of 0.9% NaCl as the control (n = 5) or carbachol (3 microg/kg, n = 5). Central carbachol did not significantly change fetal low voltage (LV) and high voltage (HV) ECoG temporal distributions. However, swallowing activity during LV ECoG was elevated significantly after intracerebroventricular carbachol. Associated with the swallowing activation, c-fos immunoreactivity in the putative dipsogenic center, subfornical organ, was enhanced significantly. The fetal urine flow rate and renal Na+, K+, and Cl(-) excretion were markedly increased following intracerebroventricular carbachol and sustained at the high level for at least 2 h. The results indicate that the central cholinergic mechanism is established and functional in regulation of fetal behavior and renal excretion at least at 0.9 gestation, which plays an important role in maintenance of fetal body fluid homeostasis.

Newly identified patterns of Pax2 expression in the developing mouse forebrain.

BACKGROUND: The availability of specific markers expressed in different regions of the developing nervous system provides a useful tool for the study of mouse mutants. One such marker, the transcription factor Pax2, is expressed at the midbrain-hindbrain boundary and in the cerebellum, spinal cord, retina, optic stalk, and optic chiasm. We recently described a group of diencephalic cells that express Pax2 as early as embryonic day (E) 10.5, and become part of the eminentia thalami by E11.5. The discovery of this previously undescribed cell population prompted us to examine Pax2 protein expression in the developing mouse forebrain in more detail. RESULTS: We determined the expression pattern of Pax2 in the forebrain of wild type mouse embryos between E10.5 and postnatal day (P) 15. Pax2 expression was detected in the septum of the basal forebrain, hypothalamus, eminentia thalami and in the subfornical organ. To evaluate Pax2 as a marker for septal cells, we examined Pax2 expression in Pax6Sey/Sey mutants, which have an enlarged septum. We found that Pax2 clearly marks a population of septal cells equivalent to that seen in wild types, indicating its utility as a marker of septal identity. These cells did not express the GABAergic marker calbindin nor the cholinergic marker choline acetyltransferase and were not detectable after P15. CONCLUSION: Pax2 is expressed in populations of cells within the developing septum, hypothalamus, and eminentia thalami. It seems especially useful as a marker of the telencephalic septum, because of its early, strong and characteristic expression in this structure. Further, its expression is maintained in the enlarged septum of Pax6Sey/Sey mutants.

Intact osmoregulatory centers in the preterm ovine fetus: Fos induction after an osmotic challenge.

We previously demonstrated a functional systemic dipsogenic response in the near-term fetal sheep (128-130 days; 145 days = full-term) with swallowing activity stimulated in response to central and systemic hypertonic saline. Preterm fetal sheep (110-115 days) do not consistently demonstrate swallowing in response to hypertonic stimuli, and it is unclear whether this is due to immaturity of osmoreceptor mechanisms or neuronal pathways activating swallowing motor neurons. To determine whether osmoreceptive regions in the preterm fetus are activated by changes in plasma tonicity, we examined Fos expression with immunostaining in these neurons in response to an osmotic challenge. Nine preterm fetal sheep [five hypertonic saline-treated fetuses (Hyp) and four isotonic saline-treated fetuses (Iso)] were prepared with vascular and intraperitoneal catheters. Seventy-five minutes before tissue collection, hypertonic (1.5 M) or isotonic saline was infused (12 ml/kg) via an intraperitoneal catheter to fetuses. Brains were examined for patterns of neuronal activation (demonstrated by Fos protein expression). Hyp demonstrated increases in plasma osmolality (~10 mosmol/kg H(2)O) and Na concentrations (5 meq/l). Increased Fos expression was detected in Hyp in the organum vasculosum of the lamina terminalis (OVLT), subfornical organ (SFO), median preoptic nucleus (MnPO), supraoptic (SON), and paraventricular nuclei (PVN) compared with Iso animals. Neuronal activation within the OVLT, SFO, and MnPO indicates intact osmoregulatory mechanisms, whereas activation of the SON and PVN suggests intact fetal neural pathways to arginine vasopressin neurons. These results suggest that preterm fetal swallowing insensitivity to osmotic stimuli may be due to immaturity of integrated motor neuron pathways.

Early development of the human area postrema and subfornical organ.

The first appearance and early development of two circumventricular organs, the area postrema (AP) and the subfornical organ (SFO), were investigated in human embryos and fetuses from the 4th to the 40th gestational weeks (GW). The AP appears very early in development, during the GW 10; its high vascularization can be seen from GW14, and differentiated neurons are observed from GW 16. The SFO is characterized by a late onset of development. It can first be distinguished at GW 17, but it does not attain cytological differentiation until the last weeks of gestation. It is suggested that the AP has important functions during fetal life, which are related to normal fetal weight and growth; in contrast the SFO, which is connected with drinking behavior and salt/water balance, seems to play a less essential role in early fetal life.

Timetables of cytogenesis in the rat subfornical organ.

Timetables of neurogenesis and ependymal cell production in the rat subfornical organ (SFO) were determined by examining the offspring of pregnant rats injected with [3H]thymidine on E13-E14, E14-E156, ... E21-E22, respectively. The proportion of postmitotic cells originating each embryonic day was determined by analyzing, in the adult offspring, the progressive reduction in the proportion of labeled precursors from the maximum amount seen in the E13-E14 group. Neurogenesis was found to occur over an extended period of time, beginning on E12 and continuing through E21. Ependymal cells were generated E15 through E21. Both neuron and ependymal cell production occurred in a triphasic pattern and followed an anterior (older) to posterior (younger) gradient. The anterior to posterior production gradient may be related to the morphological variation which exists along this plane. A production gradient intrinsic to a particular levels was found only in the posterior SFO, where peripheral neurons form earlier than core neurons. That neurogenetic gradient may be related to the core-periphery topographical patterns found in other studies, and suggests that the core neurons, since they are among the last to be formed, may be interneurons.

Acetylcholinesterase activity during the ontogenesis of the subfornical organ of the rat.

From 16 to 18 days of fetal life (fl) the subfornical organ (SFO) of the rat shows diffuse acetylcholinesterase (AChE) activity. From 21 fl its definitive pattern begins to be organized and is achieved at 30 days of postnatal life (pl). This pattern is characterized by a homogeneous AChE activity in the rostral region, a ring-like distribution in the anterior medial region and a hoof-like distribution in the posterior medial region. The caudal region shows intense, uniform activity. From the rostral region two tracts emerge, ventral and dorsal, that extend to different diencephalo-telencephalic areas. From the SFO caudal region another tract emerges that extends to posterior areas.