cFOS as a biomarker of activity maturation in the hippocampal formation.

We explored the potential for cFOS expression as a marker of functional development of "resting-state" waking activity in the extended network of the hippocampus and entorhinal cortex. We examined sleeping and awake mice at (P)ostnatal days 5, 9, 13, and 17 as well as in adulthood. We find that cFOS expression is state-dependent even at 5 days old, with reliable staining occurring only in the awake mice.

Novel localizations of TRPC5 channels suggest novel and unexplored roles: A study in the chick embryo brain.

Mammalian TRPC5 channels are predominantly expressed in the brain, where they increase intracellular Ca and induce depolarization. Because they augment presynaptic vesicle release, cause persistent neural activity, and show constitutive activity, TRPC5s could play a functional role in late developmental brain events. We used immunohistochemistry to examine TRPC5 in the chick embryo brain between 8 and 20 days of incubation, and provide the first detailed description of their distribution in birds and in the whole brain of any animal species.

Melanin-concentrating hormone (MCH) neurons in the developing chick brain.

The present study was undertaken because no previous developmental studies exist on MCH neurons in any avian species. After validating a commercially-available antibody for use in chickens, immunohistochemical examinations first detected MCH neurons around embryonic day (E) 8 in the posterior hypothalamus. This population increased thereafter, reaching a numerical maximum by E20.

Optimized CUBIC protocol for three-dimensional imaging of chicken embryos at single-cell resolution.

The CUBIC tissue-clearing protocol has been optimized to produce translucent immunostained whole chicken embryos and embryo brains. When combined with multispectral light-sheet microscopy, the validated protocol presented here provides a rapid, inexpensive and reliable method for acquiring accurate histological images that preserve three-dimensional structural relationships with single-cell resolution in whole early-stage chicken embryos and in the whole brains of late-stage embryos.

Activation of state-regulating neurochemical systems in newborn and embryonic chicks.

Coordinated activity in different sets of widely-projecting neurochemical systems characterize waking (W) and sleep (S). How and when this coordination is achieved during development is not known. We used embryos and newborns of a precocial bird species (chickens) to assess developmental activation in different neurochemical systems using cFos expression, which has been extensively employed to examine cellular activation during S and W in adult mammals.

Chick embryos have the same pattern of hypoxic lower-brain activation as fetal mammals.

cFos expression (indicating a particular kind of neuronal activation) was examined in embryonic day (E) 18 chick embryos after exposure to 4 h of either normoxia (21% O2), modest hypoxia (15% O2), or medium hypoxia (10% O2). Eight regions of the brainstem and hypothalamus were surveyed, including seven previously shown to respond to hypoxia in late-gestation mammalian fetuses (Breen et al., 1997; Nitsos and Walker, 1999b).

Early expression of hypocretin/orexin in the chick embryo brain.

Hypocretin/Orexin (H/O) neuropeptides are released by a discrete group of neurons in the vertebrate hypothalamus which play a pivotal role in the maintenance of waking behavior and brain state control. Previous studies have indicated that the H/O neuronal development differs between mammals and fish; H/O peptide-expressing cells are detectable during the earliest stages of brain morphogenesis in fish, but only towards the end of brain morphogenesis (by ∼ 85% of embryonic development) in rats. The developmental emergence of H/O neurons has never been previously described in birds.

Opposing effects of hypoxia on catecholaminergic locus coeruleus and hypocretin/orexin neurons in chick embryos.

Terrestrial vertebrate embryos face a risk of low oxygen availability (hypoxia) that is especially great during their transition to air-breathing. To better understand how fetal brains respond to hypoxia, we examined the effects of low oxygen availability on brain activity in late-stage chick embryos (day 18 out of a 21-day incubation period). Using cFos protein expression as a marker for neuronal activity, we focused on two specific, immunohistochemically identified cell groups known to play an important role in regulating adult brain states (sleep and waking): the noradrenergic neurons of the Locus Coeruleus (NA-LC), and the Hypocretin/Orexin (H/O) neurons of the hypothalamus.

Design and implementation of a wireless in-ovo EEG/EMG recorder.

The developmental origins of sleep and brain activity rhythms in higher vertebrate animals (birds and mammals) are currently unknown. In order to create an experimental system in which these could be better elucidated, we designed, built and tested a system for recording EEG and EMG signals in-ovo from chicken embryos incubated for 16-21 days. This system can remain attached to the individual subject through the process of hatching and continue to be worn post-natally.

Fos and FRA protein expression in rat precerebellar structures during the Neurolab Space Mission.

Changes in gene expression were examined in precerebellar structures during and after space flight. These structures included the inferior olive (IO), the source of climbing fibers, and the lateral reticular nucleus (LRt) and basilar pontine nuclei (PN), sources of mossy fibers. We examined two immediate early gene products with two different time courses of expression: Fos, which persists only for a few (6-8)h after activation and FRA expression, which lasts for longer periods of time, i.

Fos and FRA protein expression in rat nucleus paragigantocellularis lateralis during different space flight conditions.

The nucleus paragigantocellularis lateralis (LPGi) exerts a prominent excitatory influence over locus coeruleus (LC) neurons, which respond to gravity signals. We investigated whether adult albino rats exposed to different gravitational fields during the NASA Neurolab Mission (STS-90) showed changes in Fos and Fos-related antigen (FRA) protein expression in the LPGi and related cardiovascular, vasomotor, and respiratory areas. Fos and FRA proteins are induced rapidly by external stimuli and return to basal levels within hours (Fos) or days (FRA) after stimulation.