Figure 1.
Neuropathology in the dorsal root ganglion and enteric nervous system MPS IIIB mice.
DRG (a) and small intestine (b) isolated from 6 mo old MPS IIIB mice were processed for histopathology using toluidine blue (TB) staining, TEM and IF for LAMP-1 or GFAP. a. DRG: Top panel: TB staining,; middle panel: TEM; bottom panel: IF for LAMP-1. N: neurons; white arrowheads: satellite cells; blue arrows: Schwann cells; red arrowheads: lysosomal storage lesions; black arrows: low density material storage under TEM; green arrowheads: zebra bodies. Red fluorescence: LAMP-1-positive signal (yellow arrowheads). b. Intestine: Left panel: TB staining. Middle panel: IF for LAMP-1 (red fluorescence). Right panel: IF for GFAP (green fluorescence). *: longitudinal muscle layer; #: circular muscle layer; yello arrowheads: neurons of myenteric plexus; black arrowheads: neurons of submucosal plexus; red arrowhead: peritoneal surface. Scale bars: 20 µm.
Figure 2.
Loss of tactile and thermal nociceptive response in MPS IIIB mice.
MPS IIIB (−/−) and wt (+/+) littermates were tested for sensory functions. a. Monofilament touch test (n = 10/group, 6 mo of age): the unquantified data were analyzed using chi-square test and logistic regression with repeated measurements. b. Tail-flick assay. c. Hot plate test. 4 mo old MPS IIIB mice: n = 15/group; wt and 6 mo-old MPS IIIB mice: n = 10/group. Data are means±sd.
Figure 3.
Lysosomal storage pathology, astrocytosis and demyelination in the spinal cord of MPS IIIB mice.
Spinal cord samples from 6 mo old MPS IIIB mice were processed for paraffin sections (4 µm) and assayed by toluidine blue staining for histopathology (A), immunohistochemistry for GFAP and Iba1 (B) and transmission electron microscopy (C). A (lumbar). a-b: wt; c-h: MPS IIIB; yellow arrowheads: motor neurons (a, c, e); red arrowheads: Ependymal calls; cc: central canal (b, d); red arrow: oligodendrocyte (f); black arrow: pericyte; *: capillary (g); green arrowhead: sensory neurons (h); black arrowhead: microglia; scale bar = 20 µm. B. IF images: red arrowheads: GFAP-positive astrocytes and processes (green fluorescence). yellow arrowheads: Iba1-positive cells (red fluorescence). Scale bar = 50 µm. GFAP- or Iba1-positive cells were counted in an area of 300×430 µm on cervical, thoracic and lumbar sections from 6 mice per group. Cell counting was performed on 3 sections/level/mouse. No. cells: means±sd. C. TEM images (lumbar): black arrows: myelinated exons and synapses; *: cytoplasm of oligodendrocytes; n: nucleus of oligodendrocyte; red arrows: lysosomal storage lesions. Scale bar = 500 nm.