Unbiased stereology series#

Neurodegeneration refers to the progressive loss of nerve cells, occurring in aging and in neurodegenerative disorders, comprising AD, PD, ALS, HD, stroke, and head trauma. Apoptotic death of hippocampal and cortical neurons is responsible for the symptoms of AD death of midbrain neurons that use the neurotransmitter dopamine underlies PD HD involves the death of neurons in the striatum, which control body movements and death of lower motor neurons manifests as ALS.

Acute neurotoxicity, brain ischemia and trauma induce necrosis of a small brain area, which then propagates neuronal cell loss by apoptosis to a larger brain area. Neuronal injury or cell death refers to many acute and chronic neurological conditions, including AlzheimerвЂ™s disease (AD), ParkinsonвЂ™s disease (PD) and HuntingtonвЂ™s disease (HD), stroke, trauma, epilepsy, multiple sclerosis and amyotrophic lateral sclerosis (ALS). Neuronal injury and neurodegeneration are hallmark pathologies of a variety of acute and chronic neurological conditions such as traumatic brain injury, stroke and epilepsy. This unbiased stereology protocol allows for sensitive, medium-throughput counting of total neurons in any brain region, and thus provides a quantitative tool for studies of neuronal injury and neurodegeneration in a variety of acute brain injury and chronic neurological models. The total neuron counts were consistent with literature values from sophisticated systems, which are more expensive than our stereology system. The protocol has greater predictive power for absolute counts as it is based on 3D features rather than 2D images.

This protocol yielded an estimate of 1.5 million total neurons and 0.05 million PV(+) interneurons within the rat hippocampus. Regional tissue volume was determined by using the Cavalieri estimator, as well as cell density and cell number were determined by using the optical disector and optical fractionator.

Each region was separately analyzed with a microscope driven by the stereology software. The sections that contain the hippocampus were then delineated into five reliably predefined subregions.

Unbiased stereology series#

Every 20th section in each series of 20 sections was processed for NeuN(+) total neuron and parvalbumin(+) interneuron immunostaining. Here, we describe an optimized stereology protocol for quantification of neuronal injury and neurodegeneration by unbiased counting of neurons and interneurons. However, conventional qualitative scoring-based protocols are superficial and less reliable for use in studies of neuroprotection evaluations. Quantification of absolute neuron and interneuron counts in various brain regions is essential to understand the impact of neurological insults or neurodegenerative disease progression in animal models. Neuronal injury and neurodegeneration are the hallmark pathologies in a variety of neurological conditions such as epilepsy, stroke, traumatic brain injury, ParkinsonвЂ™s disease and AlzheimerвЂ™s disease. Department of Neuroscience and Experimental Therapeutics, Texas A&M University Health Science Center, College of Medicine, Bryan, TX, USA.Golub, Jonathan Brewer, Xin Wu, Ramkumar Kuruba, Jenessa Short, Maunica Manchi, Megan Swonke, Iyan Younus and Doodipala Samba Reddy *